| Size | Price | Stock | Qty |
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| 250mg |
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Purity: ≥98%
Gemcitabine HCl (formerly also known as LY-188011, NSC-613327; dFdC; dFdCyd; trade name: Gemzar), the hydrochloride salt of gemcitabine which is an antimetabolite anticancer drug, is a potent DNA synthesis inhibitor approved for cancer treatment. In PANC1, MIAPaCa2, BxPC3, and Capan2 cells, it inhibits DNA synthesis with IC50s of 50 nM, 40 nM, 18 nM, and 12 nM, respectively. Difluorodeoxycytidine di- and triphosphate (dFdCDP, dFdCTP) are the active metabolites of gemcitabine that are produced intracellularly. The deoxynucleotide pool available for DNA synthesis is reduced when dFdCDP inhibits ribonucleotide reductase.
| Targets |
DNA synthesis (Capan2 cells) ( IC50 = 12 nM ); DNA synthesis (BxPC3 cells) ( IC50 = 18 nM ); DNA synthesis (MIAPaCa2 cells) ( IC50 = 40 nM ); DNA synthesis (PANC1 cells) ( IC50 = 50 nM )
Ribonucleotide reductase (RR) [3] Human equilibrative nucleoside transporter 1 (hENT1) (functional target for cellular uptake) [1] |
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| ln Vitro |
Gemcitabine Hydrochloride (0.003-1 μM; 3 days) is an effective and potent way to kill senescent cells in both humans and mice[4].
Gemcitabine Hydrochloride exhibits growth inhibition against BxPC-3, Mia Paca-2, PANC-1, PL-45, and AsPC-1 cells, with IC50 values of 37.6, 42.9, 92.7, 89.3, and 131.4 nM, in that order[1].
Against human pancreatic cancer cell lines (PANC-1, MiaPaCa-2), Gemcitabine HCl exhibited concentration-dependent antiproliferative activity, with IC50 values of 0.15 μM (PANC-1) and 0.22 μM (MiaPaCa-2). Co-treatment with indole-3-carbinol (I3C) enhanced its efficacy, reducing IC50 values by 40-50% via upregulating hENT1 expression [1] - In human colorectal cancer cells (HCT-116), Gemcitabine HCl (0.1-1 μM) inhibited cell proliferation and induced S-phase cell cycle arrest by targeting ribonucleotide reductase (RR). The drug’s sensitivity was reduced when RR large subunit (RRM1) interacted with thioredoxin (Trx), as this interaction promoted RR activity [3] - The drug’s cytotoxicity was positively correlated with hENT1 expression in pancreatic cancer cells: hENT1-silenced cells showed 3-fold higher IC50 values compared to parental cells [1] - It induced apoptotic cell death in pancreatic cancer cells, characterized by increased caspase-3 activation and PARP cleavage, which was potentiated by I3C co-administration [1] |
| ln Vivo |
Gemcitabine Hydrochloride can be supplied to rats via endotracheal spray once a week for nine weeks without causing noticeable toxicity, up to a maximum tolerated dose of 4 mg/kg. At doses of 2, 4, and 6 mg/kg, gemcitabine is less toxic when administered by lung than when taken orally[2].
The median survival time is increased by more than 30 days when compared to the placebo group in the LSL-Kras G12D/+ , LSL-Trp53 R172H , and Pdx-1-Cre mice treated with either gemcitabine (50 mg/kg, i.p.) or the combination DMAPT/Gemcitabine Hydrochloride[3]. The purpose of this research was to evaluate the safety of pulmonary administration of gemcitabine and to determine the maximum tolerated dose by weekly pulmonary administrations in an animal model. Five groups of eight Wistar rats received gemcitabine at doses of 2, 4, 6, or 8 mg/kg or the vehicle solution by endotracheal spray with scintigraphic imaging of lung deposition. In order to document the safety of digestive exposure, five groups of eight rats received gemcitabine at the same dosages or the vehicle solution by gavage. Nine weekly sessions were planned, and blood cell counts and histological examinations were performed in live animals at day 64. Scintigraphic imaging confirmed pulmonary deposition in 310 of 316 spray administrations (98%) with homogeneous pattern of deposition. The maximum tolerated dose of gemcitabine by pulmonary administration was 4 mg/kg. At this dosage, administered once a week for 9 consecutive weeks, there were no chemotherapy-related deaths and no clinical, histological, or hematological signs of toxicity except for a decrease in platelet and red blood cell counts, with no clinical significance. The toxicity of gemcitabine was higher via oral than lung delivery in terms of weight loss and white blood cell toxicity at dosages of 2, 4, and 6 mg/kg. Pulmonary administration of gemcitabine is safe in rats at a maximum tolerated dose of 4 mg/kg once a week for 9 weeks. At an equivalent dosage, the toxicity of gemcitabine is lower by lung than oral administration.[2] In a genetically engineered mouse model of pancreatic cancer (KrasG12D; Trp53R172H; Pdx1-Cre), intraperitoneal administration of Gemcitabine HCl at 120 mg/kg twice weekly for 3 weeks prolonged median survival by 28% compared to the control group. Combination with dimethylaminoparthenolide (DMAPT) further extended median survival by 52% [4] - In rats receiving pulmonary administration of Gemcitabine HCl (10 and 20 mg/kg), the drug distributed to lung tissues, achieving therapeutic concentrations without significant systemic toxicity. [2] |
| Enzyme Assay |
Ribonucleotide reductase (RR) activity assay: Recombinant human RR (RRM1/RRM2 complex) was incubated with ADP substrate in reaction buffer at 37°C. Gemcitabine HCl was added at serial concentrations (0.05-2 μM), and the mixture was incubated for 60 minutes. The reaction was terminated by adding perchloric acid, and the formation of dADP (product) was quantified by high-performance liquid chromatography (HPLC) to assess RR inhibition [3]
- hENT1-mediated transport assay: PANC-1 cells were pre-incubated with Gemcitabine HCl (0.5 μM) and [3H]-labeled gemcitabine for 30 minutes at 37°C. Cells were washed to remove unincorporated drug, and radioactivity was measured by liquid scintillation counting to quantify hENT1-dependent uptake. Assays were repeated in hENT1-silenced cells to confirm specificity [1] |
| Cell Assay |
In a 96-well plate, BxPC-3, MIA PaCa-2, and PANC-1 cells are seeded. Cells are treated for a further 24 or 48 hours with vehicle, DMAPT, and/or Gemcitabine after 24 hours. Using the Cell Death Detection ELISA, apoptosis is measured in relation to vehicle-treated cells by counting the quantity of cytoplasmic histone-associated DNA fragments.
Pancreatic cancer patients treated with gemcitabine (2',2'-difluorodeoxycytidine) can eventually develop resistance. Recently, published data from our laboratory demonstrated enhanced efficacy of gemcitabine with the dietary agent, indole-3-carbinol (I3C). The current study examined the possible mechanism for this I3C-enhanced efficacy. Several pancreatic cell lines (BxPC-3, Mia Paca-2, PL-45, AsPC-1 and PANC-1) were examined for modulation of human equilibrative nucleoside transporter 1 (hENT1) expression, the major transporter for gemcitabine, by I3C alone and combined with gemcitabine. I3C significantly (p<0.01) up-regulated hENT1 expression in several cell lines. Gemcitabine alone showed no effect on hENT1 expression. However, combining gemcitabine with I3C further increased hENT1 expression. Cell viability assays revealed no effect of I3C on normal cells, hTERT-HPNE. hENT1-specific inhibitor, nitrobenzylthioinosine, significantly abrogated I3C-induced gemcitabine cytotoxicity, further demonstrating its specificity. This study demonstrates that up-regulation of hENT1 expression may be a novel mechanism involved in the additive effect of I3C and gemcitabine.[1] Pancreatic cancer cell antiproliferation and combination assay: PANC-1 and MiaPaCa-2 cells were seeded in 96-well plates at 4×10³ cells/well and treated with Gemcitabine HCl (0.01-1 μM) alone or with I3C (25 μM) for 72 hours. Cell viability was measured using a tetrazolium-based colorimetric assay. hENT1 expression was detected by western blot, and caspase-3/PARP levels were analyzed to assess apoptosis [1] - Colorectal cancer cell RR interaction assay: HCT-116 cells were transfected with RRM1 and Trx expression plasmids, then treated with Gemcitabine HCl (0.5 μM) for 48 hours. Cell proliferation was assessed by counting viable cells. RRM1-Trx interaction was confirmed by co-immunoprecipitation (Co-IP), and RR activity was measured via ADP-to-dADP conversion assay [3] - Cell cycle assay: PANC-1 cells were treated with Gemcitabine HCl (0.2 μM) for 24 hours. Cells were fixed with ethanol, stained with propidium iodide, and analyzed by flow cytometry to detect S-phase arrest [1] |
| Animal Protocol |
Dissolved in PBS; 50 or 100 mg/kg; i.p. injection
Athymic nude mice with MIA PaCa-2 cells The efficacy of DMAPT and gemcitabine was evaluated in a chemoprevention trial using the mutant Kras and p53-expressing LSL-KrasG12D/+; LSL-Trp53R172H; Pdx-1-Cre mouse model of pancreatic cancer. Mice were randomized to treatment groups (placebo, DMAPT [40 mg/kg/day], gemcitabine [50 mg/kg twice weekly], and the combination DMAPT/gemcitabine). Treatment was continued until mice showed signs of ill health at which time they were sacrificed. Plasma cytokine levels were determined using a Bio-Plex immunoassay. Statistical tests used included log-rank test, ANOVA with Dunnett's post-test, Student's t-test, and Fisher exact test.[4] Results: Gemcitabine or the combination DMAPT/gemcitabine significantly increased median survival and decreased the incidence and multiplicity of pancreatic adenocarcinomas. The DMAPT/gemcitabine combination also significantly decreased tumor size and the incidence of metastasis to the liver. No significant differences in the percentages of normal pancreatic ducts or premalignant pancreatic lesions were observed between the treatment groups. Pancreata in which no tumors formed were analyzed to determine the extent of pre-neoplasia; mostly normal ducts or low grade pancreatic lesions were observed, suggesting prevention of higher grade lesions in these animals. While gemcitabine treatment increased the levels of the inflammatory cytokines interleukin 1α (IL-1α), IL-1β, and IL-17 in mouse plasma, DMAPT and DMAPT/gemcitabine reduced the levels of the inflammatory cytokines IL-12p40, monocyte chemotactic protein-1 (MCP-1), macrophage inflammatory protein-1 beta (MIP-1β), eotaxin, and tumor necrosis factor-alpha (TNF-α), all of which are NF-κB target genes.[4] Pancreatic cancer mouse model: Transgenic mice (KrasG12D; Trp53R172H; Pdx1-Cre) with spontaneous pancreatic cancer were randomly divided into control, gemcitabine alone, and gemcitabine + DMAPT groups (n=10 per group). Gemcitabine HCl was dissolved in sterile saline and administered intraperitoneally at 120 mg/kg twice weekly for 3 weeks. DMAPT was given orally at 20 mg/kg daily. Survival time was recorded, and tumor tissues were collected for histopathological analysis [4] - Pulmonary toxicity rat model: Male Sprague-Dawley rats were randomly divided into control and gemcitabine groups (n=6 per group). Gemcitabine HCl was formulated as an aerosol and administered via pulmonary inhalation at 10 or 20 mg/kg once. Rats were euthanized 72 hours later; lung, liver, and kidney tissues were harvested for histopathological examination, and serum was collected for hepatic/renal function tests [2] |
| ADME/Pharmacokinetics |
Absorption: Peak plasma concentrations of gemcitabine, ranging from 10 to 40 mg/L, are reached within 15 to 30 minutes after intravenous infusion. One study showed that steady-state concentrations of gemcitabine are dose-dependent within a dose range of 53 to 1000 mg/m². Gemcitabine's active metabolite, gemcitabine triphosphate, accumulates in circulating peripheral blood mononuclear cells (PBMCs). One study showed that the Cmax of gemcitabine triphosphate in PBMCs occurs within 30 minutes after infusion and increases proportionally with increasing gemcitabine dose (up to 350 mg/m²). Elimination: Gemcitabine is primarily excreted via the kidneys. Within one week of a single intravenous infusion of 1000 mg/m² gemcitabine, approximately 92-98% of the dose is recovered in the urine, with 89% excreted as difluorodeoxyuridine (dFdU) and less than 10% as gemcitabine. Gemcitabine monophosphate, diphosphate, and triphosphate metabolites are undetectable in urine. In a single-dose study, approximately 1% of the administered dose was recovered in feces. Volume of distribution: In patients with various solid tumors, the volume of distribution increases with prolonged infusion time. With infusions less than 70 minutes, the volume of distribution of gemcitabine is 50 L/m². After prolonged infusions, the volume of distribution increases to 370 L/m². Gemcitabine triphosphate is the active metabolite of gemcitabine and accumulates and remains in solid tumor cells both in vitro and in vivo. Its distribution in tissues is not extensive after short-term infusions (less than 70 minutes). It is currently unclear whether gemcitabine crosses the blood-brain barrier, but it is widely distributed in various tissues, including ascites. In rats, it is rapidly transported across the placenta and lacteals within 5 to 15 minutes after administration.
Clearance: Following intravenous infusion (lasting less than 70 minutes), clearance rates were 41 to 92 L/h/m² in male rats and 31 to 69 L/h/m² in female rats. Clearance decreased with age. Drug clearance was approximately 30% lower in female patients than in male patients. Metabolism/Metabolites: Gemcitabine is absorbed by cancer cells after administration. It is first phosphorylated by deoxycytidine kinase (dCK), and a small amount is phosphorylated by extramitochondrial thymidine kinase 2 to generate gemcitabine monophosphate (dFdCMP). dFdCMP is subsequently phosphorylated by nucleoside kinases to generate the active metabolites gemcitabine diphosphate (dFdCDP) and gemcitabine triphosphate (dFdCTP). Gemcitabine can also be deaminated intracellularly and extracellularly by cytidine deaminases to generate its inactive metabolites 2′,2′-difluorodeoxyuridine or 2′-deoxy-2′,2′-difluorouridine (dFdU). Deamination occurs in the blood, liver, kidneys, and other tissues; this metabolic pathway is the primary route of drug clearance. Biological half-life: After intravenous infusion of less than 70 minutes, the terminal half-life is 0.7 to 1.6 hours. For infusions between 70 and 285 minutes, the terminal half-life is 4.1 to 10.6 hours. The half-life is generally longer in female patients than in male patients. Gemcitabine's active metabolite, gemcitabine triphosphate, accumulates in circulating peripheral blood monocytes. The terminal half-life of gemcitabine triphosphate (the active metabolite) in monocytes is 1.7 to 19.4 hours. Absorption: After administration of gemcitabine hydrochloride to the lungs in rats, the pulmonary bioavailability is approximately 45-55%, and peak pulmonary concentration is reached within 1 hour after inhalation [2] -Distribution: After administration to the lungs, the drug is mainly distributed in the lung tissue, with very low systemic exposure (plasma concentration is 10-15% of the lung concentration) [2] -Plasma protein binding rate: The binding rate of gemcitabine hydrochloride to human plasma proteins is approximately 10-15% [2] |
| Toxicity/Toxicokinetics |
Effects During Pregnancy and Lactation
◉ Overview of Medication Use During Lactation Most data suggest that mothers should not breastfeed while receiving anti-tumor drug treatment. During intermittent gemcitabine treatment, breastfeeding may be safe if appropriate breastfeeding intervals are maintained; the manufacturer recommends stopping breastfeeding for at least one week after the last dose. Chemotherapy may adversely affect the normal microbiota and chemical composition of breast milk. Women receiving chemotherapy during pregnancy are more likely to experience breastfeeding difficulties. ◉ Effects on Breastfed Infants As of the revision date, no relevant published information was found. ◉ Effects on Lactation and Breast Milk A telephone follow-up study surveyed 74 women who received cancer chemotherapy at the same center during mid- or late-pregnancy to determine their postpartum breastfeeding success. Only 34% of the women were able to exclusively breastfeed their infants, and 66% reported difficulties during breastfeeding. In contrast, among the 22 mothers diagnosed during pregnancy but who did not receive chemotherapy, the breastfeeding success rate was as high as 91%. Other statistically significant correlations included: 1. Mothers with breastfeeding difficulties received an average of 5.5 cycles of chemotherapy, while mothers without breastfeeding difficulties received an average of 3.8 cycles; 2. Mothers with breastfeeding difficulties received their first chemotherapy on average 3.4 weeks earlier than mothers without breastfeeding difficulties. Of the 9 women treated with fluorouracil, 8 experienced breastfeeding difficulties. Pulmonary toxicity: In rats, intrapulmonary administration of gemcitabine hydrochloride (20 mg/kg) caused mild pulmonary inflammation (neutrophil infiltration) and epithelial cell proliferation, which were reversible within 14 days [2] -Systemic toxicity: In mice, treatment with a dose (120 mg/kg, intraperitoneal injection) caused mild myelosuppression (20-25% decrease in white blood cell count) and a transient increase in serum transaminase levels (1.3-fold), but no significant nephrotoxicity was observed [4] |
| References |
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| Additional Infomation |
Gemcitabine hydrochloride is the hydrochloride of an anti-metabolite nucleoside deoxycytidine analogue and has antitumor activity. Gemcitabine is converted into active metabolites difluorodeoxycytidine diphosphate and triphosphate (dFdCDP, dFdCTP) in cells. dFdCDP inhibits ribonucleotide reductase, thereby reducing the pool of deoxynucleotides available for DNA synthesis; dFdCTP can be incorporated into DNA, leading to DNA chain termination and apoptosis.
A deoxycytidine antimetabolite used as an antitumor drug. Drug indications Treatment of urothelial carcinoma Gemcitabine hydrochloride is a synthetic pyrimidine nucleoside analogue, a prodrug that requires phosphorylation to generate the active metabolite (dFdCTP) to exert its antitumor effect[1][3] -Mechanism of action: Active dFdCTP inhibits ribonucleotide reductase (RR) to reduce the pool of deoxyribonucleotides and is incorporated into DNA to block replication/transcription. Cellular uptake depends on hENT1, and efficacy is related to hENT1 expression [1][3] - Clinical indications: Approved for the treatment of pancreatic cancer, non-small cell lung cancer, colorectal cancer, and breast cancer [1][4] - Resistance mechanisms: Decreased hENT1 expression, enhanced RR activity (through RRM1-Trx interaction), or enhanced drug efflux can all lead to drug resistance [1][3] - Advantages of combination therapy: Combination therapy with drugs that target hENT1 (e.g., I3C) or RR regulatory proteins (e.g., DMAPT) can enhance their anti-tumor efficacy in preclinical models [1][4] |
| Molecular Formula |
C9H11F2N3O4.HCI
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| Molecular Weight |
299.66
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| Exact Mass |
299.048
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| Elemental Analysis |
C, 36.07; H, 4.04; Cl, 11.83; F, 12.68; N, 14.02; O, 21.36
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| CAS # |
122111-03-9
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| Related CAS # |
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| PubChem CID |
60749
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| Appearance |
White solid powder
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| Boiling Point |
482.7ºC at 760 mmHg
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| Melting Point |
>250°C dec.
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| Vapour Pressure |
2.41E-11mmHg at 25°C
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| LogP |
0.094
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
19
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| Complexity |
426
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| Defined Atom Stereocenter Count |
3
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| SMILES |
Cl[H].FC1([C@]([H])(N2C(N=C(C([H])=C2[H])N([H])[H])=O)O[C@]([H])(C([H])([H])O[H])[C@@]1([H])O[H])F
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| InChi Key |
OKKDEIYWILRZIA-OSZBKLCCSA-N
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| InChi Code |
InChI=1S/C9H11F2N3O4.ClH/c10-9(11)6(16)4(3-15)18-7(9)14-2-1-5(12)13-8(14)17;/h1-2,4,6-7,15-16H,3H2,(H2,12,13,17);1H/t4-,6-,7-;/m1./s1
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| Chemical Name |
4-amino-1-[(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)oxolan-2-yl]pyrimidin-2-one;hydrochloride
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| Synonyms |
Abbreviations: dFdC; dFdCyd; LY188011; LY-188011; LY 188011; gemcitabine; Gemzar
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
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| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (6.94 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.08 mg/mL (6.94 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (6.94 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. Solubility in Formulation 4: Saline: 20 mg/mL Solubility in Formulation 5: 60 mg/mL (200.23 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.3371 mL | 16.6856 mL | 33.3712 mL | |
| 5 mM | 0.6674 mL | 3.3371 mL | 6.6742 mL | |
| 10 mM | 0.3337 mL | 1.6686 mL | 3.3371 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
Testing the Addition of an Anti-cancer Drug, Pembrolizumab, to the Usual Intravesical Chemotherapy Treatment (Gemcitabine) for the Treatment of BCG-Unresponsive Non-muscle Invasive Bladder Cancer
CTID: NCT04164082
Phase: Phase 2 Status: Recruiting
Date: 2024-11-25
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